Comparison of fluorescence and light scattering based methods to assess formation and stability of protein-protein complexes.
J Struct Biol
; 175(2): 216-23, 2011 Aug.
Article
em En
| MEDLINE
| ID: mdl-21536135
ABSTRACT
Thermal shift methods such as differential scanning fluorimetry and differential static light scattering are widely used to identify stabilizing conditions for proteins that might promote crystallization. Here we report a comparison of the two methods when applied to optimization of buffer conditions for protein-protein complexes. Most of the protein complexes under study were amenable to analysis using these two techniques. Protein complexes behave towards thermal denaturation in a manner similar to single proteins, showing a more or less sharp transition consistent with a two-state model of unfolding. A comparison of the melting and aggregation temperatures for single components and the reconstituted complexes can provide additional evidence for complex formation and can be used to identify buffer conditions in which protein-protein complex formation is favored.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Conformação Proteica
/
Espalhamento de Radiação
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Complexos Multiproteicos
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Multimerização Proteica
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Fluorometria
Tipo de estudo:
Prognostic_studies
Idioma:
En
Revista:
J Struct Biol
Ano de publicação:
2011
Tipo de documento:
Article